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发形霞水母触手提取物的蛋白稳定性及其溶血活性

常银龙,  肖良,  郑杰民,  王倩倩,  张黎明

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常银龙, 肖良, 郑杰民, 王倩倩, 张黎明. 发形霞水母触手提取物的蛋白稳定性及其溶血活性[J]. 药学实践与服务, 2014, 32(6): 434-439. doi: 10.3969/j.issn.1006-0111.2014.06.009
引用本文: 常银龙, 肖良, 郑杰民, 王倩倩, 张黎明. 发形霞水母触手提取物的蛋白稳定性及其溶血活性[J]. 药学实践与服务, 2014, 32(6): 434-439. doi: 10.3969/j.issn.1006-0111.2014.06.009
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CHANG Yinlong, XIAO liang, ZHENG Jiemin, WANG Qianqian, ZHANG Liming. Protein stability and hemolytic activity of tentacle extract from the jellyfish Cyanea capillata[J]. Journal of Pharmaceutical Practice and Service, 2014, 32(6): 434-439. doi: 10.3969/j.issn.1006-0111.2014.06.009
Citation: CHANG Yinlong, XIAO liang, ZHENG Jiemin, WANG Qianqian, ZHANG Liming. Protein stability and hemolytic activity of tentacle extract from the jellyfish Cyanea capillata[J]. Journal of Pharmaceutical Practice and Service, 2014, 32(6): 434-439. doi: 10.3969/j.issn.1006-0111.2014.06.009
shu

发形霞水母触手提取物的蛋白稳定性及其溶血活性

doi: 10.3969/j.issn.1006-0111.2014.06.009

  • 第二军医大学海医系海洋生物技术教研室, 上海 200433

基金项目: 国家自然科学基金(41176126;81370833).

Protein stability and hemolytic activity of tentacle extract from the jellyfish Cyanea capillata

  • Department of Marine Biotechnology, Faculty of Naval Medicine, Second Military Medical University, Shanghai 200433, China

  • 摘要: 目的 探讨不同环境因素和处理方法对发形霞水母(Cyanea capillata)触手提取物(tentacle extract, TE)蛋白稳定性及其溶血活性的影响。 方法 结合蛋白浓度测定、溶血活性检测和SDS-PAGE分析等方法研究不同环境因素和处理方法对TE蛋白稳定性及其溶血活性的影响。 结果 TE溶血活性呈明显的剂量依赖关系,其半数溶血分数HU50=226 μg/ml;40℃水浴1 h,能去除TE中大量杂蛋白,但仍保持显著溶血活性;TE在4℃放置28 d,其溶血活性保持稳定,25℃下3 d内其溶血活性变化不明显;pH值对TE溶血活性的影响呈钟形曲线,在pH 6.0~11.0之间活性相对稳定,pH 8.0是TE保持溶血活性的最优pH条件;不同缓冲液对TE稳定性及其溶血活性影响显著,浓度大于26%的硫酸铵溶液对TE溶血蛋白具有良好的盐析作用。 结论 40℃水浴1 h预处理可显著减少TE中非活性蛋白组分,并降低样品黏性;4℃和pH 8.0是TE保持蛋白稳定性和溶血活性的最适条件;浓度大于26%的硫酸铵盐析有利于溶血蛋白组分富集。
    Abstract: Objective To investigate the influencing factors of the protein stability and hemolytic activity of tentacle extract (TE) from the jellyfish Cyanea capillata. Methods Effects of various factors and treatments on the protein stability and hemolytic activity of TE were explored by protein detection, hemolytic assay and SDS-PAGE analysis. Results TE caused a significant and dose-dependent hemolytic effect, and the HU50 of TE against 0.5% erythrocyte suspensions from SD rats was 226 μg/ml. A 40℃ water bath for 1 hour could effectively remove the contaminating proteins in TE. TE retained hemolytic activity at 4℃ for 28 days but it was unstable when kept at 25℃ over 3 days. TE was active in the range from pH 6.0 to 11.0 and the optimum pH was 8.0. Various buffer solutions had significantly different effects on the stability and hemolytic activity of TE, and a good salting-out effect was observed on the hemolytic protein of TE while the concentration of ammonium sulfate solutions was greater than 26%. Conclusion A 40℃ water bath for 1 hour could effectively remove the contaminating proteins in TE and reduce its viscosity. The optimum conditions for maintaining stability and hemolytic activity of TE were 4℃ and pH 8.0. The salting-out effect from 26% and more ammonium sulfate solutions would be conducive to the enrichment of hemolytic protein.
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    [2] Kang C, Munawir A, Cha M, et al. Cytotoxicity and hemolytic activity of jellyfish Nemopilema nomurai(Scyphozoa: Rhizostomeae)venom[J]. Comp Biochem Physiol C Toxicol Pharmacol, 2009, 150(1): 85-90.
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  • 收稿日期:  2013-11-25
  • 修回日期:  2014-07-07

发形霞水母触手提取物的蛋白稳定性及其溶血活性

doi: 10.3969/j.issn.1006-0111.2014.06.009
  • 第二军医大学海医系海洋生物技术教研室, 上海 200433
    • 基金项目:  国家自然科学基金(41176126;81370833).
  • 收稿日期: 2013-11-25
  • 修回日期: 2014-07-07

摘要: 目的 探讨不同环境因素和处理方法对发形霞水母(Cyanea capillata)触手提取物(tentacle extract, TE)蛋白稳定性及其溶血活性的影响。 方法 结合蛋白浓度测定、溶血活性检测和SDS-PAGE分析等方法研究不同环境因素和处理方法对TE蛋白稳定性及其溶血活性的影响。 结果 TE溶血活性呈明显的剂量依赖关系,其半数溶血分数HU50=226 μg/ml;40℃水浴1 h,能去除TE中大量杂蛋白,但仍保持显著溶血活性;TE在4℃放置28 d,其溶血活性保持稳定,25℃下3 d内其溶血活性变化不明显;pH值对TE溶血活性的影响呈钟形曲线,在pH 6.0~11.0之间活性相对稳定,pH 8.0是TE保持溶血活性的最优pH条件;不同缓冲液对TE稳定性及其溶血活性影响显著,浓度大于26%的硫酸铵溶液对TE溶血蛋白具有良好的盐析作用。 结论 40℃水浴1 h预处理可显著减少TE中非活性蛋白组分,并降低样品黏性;4℃和pH 8.0是TE保持蛋白稳定性和溶血活性的最适条件;浓度大于26%的硫酸铵盐析有利于溶血蛋白组分富集。

English Abstract

常银龙, 肖良, 郑杰民, 王倩倩, 张黎明. 发形霞水母触手提取物的蛋白稳定性及其溶血活性[J]. 药学实践与服务, 2014, 32(6): 434-439. doi: 10.3969/j.issn.1006-0111.2014.06.009
引用本文: 常银龙, 肖良, 郑杰民, 王倩倩, 张黎明. 发形霞水母触手提取物的蛋白稳定性及其溶血活性[J]. 药学实践与服务, 2014, 32(6): 434-439. doi: 10.3969/j.issn.1006-0111.2014.06.009
shu
CHANG Yinlong, XIAO liang, ZHENG Jiemin, WANG Qianqian, ZHANG Liming. Protein stability and hemolytic activity of tentacle extract from the jellyfish Cyanea capillata[J]. Journal of Pharmaceutical Practice and Service, 2014, 32(6): 434-439. doi: 10.3969/j.issn.1006-0111.2014.06.009
Citation: CHANG Yinlong, XIAO liang, ZHENG Jiemin, WANG Qianqian, ZHANG Liming. Protein stability and hemolytic activity of tentacle extract from the jellyfish Cyanea capillata[J]. Journal of Pharmaceutical Practice and Service, 2014, 32(6): 434-439. doi: 10.3969/j.issn.1006-0111.2014.06.009
shu

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